LOCUS pTrans_9xD3B_CLu 7384 bp DNA circular UNA 06-JUL-2015 DEFINITION CRISPR-Display Cypridina Luciferase-2A-Venus YFP "normalizer," transient format. ACCESSION urn.local...1435876244590.78 VERSION urn.local...1435876244590.78 KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers gene complement(1..861) /gene="bla" /label="bla gene" CDS complement(1..861) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" /label="bla CDS" sig_peptide complement(793..861) /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein." /label="bla signal peptide" -10_signal complement(905..910) /note="bla promoter (counter-clockwise); GAGACA; transcript start 2548 (complementary strand)" /label="-10 signal" -35_signal complement(926..931) /note="bla promoter (counter-clockwise); TTCAAA" /label="-35 signal" source order(1062..>1473,<6193..1061) /organism="Cloning vector pNEB193" /mol_type="other DNA" /label="source Cloning vector pNEB193" primer_bind 1413..1436 /created_by="User" /label="M13F(-47) From Genewiz" primer_bind 1419..1436 /created_by="User" /label="M13(-41)F From Genewiz" primer_bind 1439..1456 /created_by="User" /label="M13F(-21) From Genewiz" misc_feature 1493..1512 /created_by="User" /label="D3B_gRNA_Binding_Site" misc_feature 1526..1545 /created_by="User" /label="D3B_gRNA_Binding_Site" misc_feature 1564..1583 /created_by="User" /label="D3B_gRNA_Binding_Site" misc_feature 1597..1616 /created_by="User" /label="D3B_gRNA_Binding_Site" misc_feature 1640..1659 /created_by="User" /label="D3B_gRNA_Binding_Site" misc_feature 1673..1692 /created_by="User" /label="D3B_gRNA_Binding_Site" misc_feature 1711..1730 /created_by="User" /label="D3B_gRNA_Binding_Site" misc_feature 1744..1763 /created_by="User" /label="D3B_gRNA_Binding_Site" misc_feature 1777..1796 /created_by="User" /label="D3B_gRNA_Binding_Site" promoter 1821..1880 /gene="minCMV" /modified_by="User" /label="minCMVPro" misc_feature 1898..1907 /created_by="User" /modified_by="User" /label="Kozak_Sequence" sig_peptide 1904..1957 /created_by="User" /label="CLuc Signal Peptide" CDS 1904..3562 /created_by="User" /modified_by="User" /label="Cypridina_Luciferase" CDS 3563..3577 /created_by="User" /modified_by="User" /label="Linker" CDS 3578..3640 /created_by="User" /modified_by="User" /label="2A_Self-Cleaving_Peptide" CDS 3641..3646 /created_by="User" /modified_by="User" /label="Linker" CDS 3647..4363 /created_by="User" /modified_by="User" /label="Venus YFP" misc_feature complement(3693..3713) /created_by="User" /label="EGFP-N Prtimer from Genewiz" /note="Geneious type: primer_bind_reverse" primer_bind 4300..4321 /created_by="User" /label="EGFP Primer from Genewiz" CDS 4364..4372 /created_by="User" /label="Linker" CDS 4373..4429 /created_by="User" /modified_by="User" /label="P2A_Self_Cleaving_Peptide" CDS 4430..4435 /created_by="User" /label="Linker" CDS 4436..5035 /created_by="User" /label="Puromycin_Resistance_Gene" misc_feature 5057..5654 /created_by="User" /modified_by="User" /label="WPRE" terminator 5755..5989 /created_by="User" /modified_by="User" /label="deltaU3/3' LTR" misc_feature 5755..5808 /created_by="User" /modified_by="User" /label="deltaU3" LTR 5809..5989 /created_by="User" /label="3' LTR" polyA_signal 6061..6192 /created_by="User" /label="SV40 PA signal" misc_feature complement(6098..6117) /created_by="User" /label="SV40 pA Primer from Genewiz" /note="Geneious type: primer_bind_reverse" misc_feature complement(6231..6247) /created_by="User" /label="M13Reverse_Primer" /note="Geneious type: primer_bind_reverse" -10_signal complement(6280..6285) /note="Plac promoter (drives lacZalpha; counter-clockwise); TATGTT" /label="-10 signal" -35_signal complement(6304..6309) /note="Plac promoter (drives lacZalpha; counter-clockwise); TTTACA" /label="-35 signal" protein_bind complement(6329..6341) /note="CAP protein binding site" /bound_moiety="catabolite activator protein (CAP)" /label="CAP protein binding site protein bind" rep_origin complement(6626..7214) /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" /label="rep origin" misc_RNA complement(6626..7178) /note="RNAII transcript (complementary strand)" /label="RNA" -35_signal 7032..7037 /note="RNAI promoter (clockwise); TTGAAG" /label="RNAI promoter (clockwise); TTGAAG -35 signal" -10_signal 7054..7059 /note="RNAI promoter (clockwise); GCTACA" /label="RNAI promoter (clockwise); GCTACA -10 signal" misc_RNA 7068..7175 /note="RNAI transcript" /label="RNAI transcript" -10_signal complement(7188..7193) /note="RNAII promoter (counter-clockwise); CGTAAT" /label="-10 signal" -35_signal complement(7209..7214) /note="RNAII promoter (counter-clockwise); TTGAGA" /label="-35 signal" ORIGIN 1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 1021 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga 1081 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga 1141 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 1201 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat 1261 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg 1321 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 1381 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 1441 taaaacgacg gccagtgaat tcgagctcgg taccttctcg agctttgcgg taatctagat 1501 cgcccgtccc ctaggcaaag ctctaatcta gatcgcccgt cccctaggaa aggaagcagc 1561 cacatctaga tcgcccgtcc cctaggataa cgttagatct agatcgcccg tcccctaggg 1621 tgaagtagtc tttgcggtaa tctagatcgc ccgtccccta ggcaaagctc taatctagat 1681 cgcccgtccc ctaggaaagg aagcagccac atctagatcg cccgtcccct aggataacgt 1741 tagatctaga tcgcccgtcc cctagggtga agtagtatct agatcgcccg tcccctaggc 1801 cggatccacg tatgtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt 1861 ttagtgaacc gtcagatcgc ctggaggtac cgccaccgcc accatgaaga ccttaattct 1921 tgccgttgca ttagtctact gcgccactgt tcattgccag gactgtcctt acgaacctga 1981 tccaccaaac acagttccaa cttcctgtga agctaaagaa ggagaatgta ttgatagcag 2041 ctgtggcacc tgcacgagag acatactatc agatggactg tgtgaaaata aaccaggaaa 2101 aacatgttgc cgaatgtgtc agtatgtaat tgaatgcaga gtagaggccg caggatggtt 2161 tagaacattc tatggaaaga gattccagtt ccaggaacct ggtacatacg tgttgggtca 2221 aggaaccaag ggcggcgact ggaaggtgtc catcaccctg gagaacctgg atggaaccaa 2281 gggggctgtg ctgaccaaga caagactgga agtggctgga gacatcattg acatcgctca 2341 agctactgag aatcccatca ctgtaaacgg tggagctgac cctatcatcg ccaacccgta 2401 caccatcggc gaggtcacca tcgctgttgt tgagatgcca ggcttcaaca tcaccgtcat 2461 tgagttcttc aaactgatcg tgatcgacat cctcggagga agatctgtaa gaatcgcccc 2521 agacacagca aacaaaggaa tgatctctgg cctctgtgga gatcttaaaa tgatggaaga 2581 tacagacttc acttcagatc cagaacaact cgctattcag cctaagatca accaggagtt 2641 tgacggttgt ccactctatg gaaatcctga tgacgttgca tactgcaaag gtcttctgga 2701 gccgtacaag gacagctgcc gcaaccccat caacttctac tactacacca tctcctgcgc 2761 cttcgcccgc tgtatgggtg gagacgagcg agcctcacac gtgctgcttg actacaggga 2821 gacgtgcgct gctcccgaaa ctagaggaac ctgcgttttg tctggacata ctttctacga 2881 tacatttgac aaagcaagat accaattcca gggtccctgc aaggagattc ttatggccgc 2941 cgactgtttc tggaacactt gggatgtgaa ggtttcacac aggaatgttg actcttacac 3001 tgaagtagag aaagtacgaa tcaggaaaca atcgactgta gtagaactca ttgttgatgg 3061 aaaacagatt ctggttggag gagaagccgt gtccgtcccg tacagctctc agaacacttc 3121 catctactgg caagatggtg acatactgac tacagccatc ctacctgaag ctctggtggt 3181 caagttcaac ttcaagcaac tgctcgtcgt acatattaga gatccattcg atggtaagac 3241 ttgcggtatt tgcggtaact acaaccagga tttcagtgat gattcttttg atgctgaagg 3301 agcctgtgat ctgaccccca acccaccggg atgcaccgaa gaacagaaac ctgaagctga 3361 acgactctgc aatagtctct tcgccggtca aagtgatctt gatcagaaat gtaacgtgtg 3421 ccacaagcct gaccgtgtcg aacgatgcat gtacgagtat tgcctgaggg gacaacaggg 3481 tttctgtgac cacgcatggg agttcaagaa agaatgctac ataaagcatg gagacaccct 3541 agaagtacca gatgaatgca aaggcagcac cagcggcggc agtggagagg gcagaggaag 3601 tctgctaaca tgcggtgacg tcgaggagaa tcctggccca ggcagtatgg tgagcaaggg 3661 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 3721 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 3781 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 3841 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt 3901 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 3961 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 4021 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 4081 ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa 4141 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca 4201 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca 4261 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 4321 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcagcg gtgctacgaa 4381 cttcagtctc ctgaaacagg ctggggatgt ggaggagaac cccggcccag gcagcatgac 4441 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac 4501 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca 4561 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg 4621 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt 4681 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg 4741 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc 4801 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc 4861 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac 4921 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt 4981 cgagtgcccg aaggaccgcg cgacctggtg catgacccgc aagcccggtg cctagtaacg 5041 taccggttag taatgatcga caatcaacct ctggattaca aaatttgtga aagattgact 5101 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 5161 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 5221 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 5281 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 5341 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 5401 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 5461 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 5521 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 5581 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 5641 gcctccccgc ctggcgatgg taccgctaag cacaattcga gctcggtacc tttaagacca 5701 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa 5761 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg 5821 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct 5881 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt 5941 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca 6001 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg 6061 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 6121 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 6181 tatcatgtct gggtttaaac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg 6241 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 6301 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 6361 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 6421 gcggggagag gcggtttgcg tattgggccg cttcctcgct cactgactcg ctgcgctcgg 6481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 6541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 6601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 6661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 6721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 6781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 6841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 6901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 6961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 7021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 7081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 7141 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 7201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 7261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 7321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 7381 acag //